Rapid Preparation of Unsheathed Bacterial Flagella

Materials and Reagents

1. Bacterial strains (S. Typhimurium)
   
2. Sodium chloride (NaCl) (Sigma-Aldrich, catalog number: S7653 )
   
3. 100% trichloroacetic acid (Sigma-Aldrich, catalog number: T6399 )
   
4. β-Mercaptoethanol (Sigma-Aldrich, catalog number: M6250 )
   
5. Trizma^® base (powder) (Sigma-Aldrich, catalog number: T1503 )
   
6. 1 M HCl solution (Sigma-Aldrich, catalog number: H1758 )
   
7. Glycerol 1 L (Sigma-Aldrich, catalog number: 49781 )
   
8. Sodium dodecyl sulfate - 500 g (Sigma, catalog number: L4390 )
   
9. Tryptone (BD-catalog number: 211705 )
   
10. Yeast extract (BD, catalog number: 212750 )
    
11. Agar (BD, catalog number: 281230 )
    
12. Bromophenol blue (Shelton scientific, catalog number: IB74040 )
    
13. Glycerol stock for one vial (see Recipes)
    
14. LB medium (see Recipes)
    
15. 1 M Tris (see Recipes)
    
16. 2x SDS sample buffer (see Recipes)
    

Equipment

1. Falcon tubes (50 ml and 15 ml) (TPP, catalog numbers: 91050 and 91115 )
   
2. Filter Flasks (pore size 0.22 μm diameter) (TPP, catalog number: 99500 )
   
3. Eppendorf tubes (1.5 ml) (Sarstedt, catalog number: 72.690.001 )
   
4. Incubator (adjusted to 37 °C) (Memmert, model: ULE500 )
   
5. Rotary shaker (HTINFORS, model: Minitron )
   
6. Spectrophotometer (Hitachi, model: U1100 )
   
7. Thermoblock (PEQLAB, model: 1202 )
   
8. Vortex mixer (VWR International, model: 1719 )
   
9. Luer lock tips (BD PLASTIKPAK, catalog number: 302187 )
   
10. Needle (0.51 mm outer diameter) (BD Microlance sterile, model25 g x 5/8” x 100)
    
11. Sterile petri dishes (90 mm) (Sartorius, catalog number: 14-555-735 )
    
12. Centrifuge (HERAEUS, model: PICO17 )
    
13. Microcentrifuge (NeoLab, model: microcentrifuge II )
    
14. pH meter (Hanna instruments, model: pH211 )
    

Procedure